CN104193988A - Method for flocculating and sterilizing fermentation solution of epsilon-polylysine - Google Patents
Method for flocculating and sterilizing fermentation solution of epsilon-polylysine Download PDFInfo
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- CN104193988A CN104193988A CN201410441173.7A CN201410441173A CN104193988A CN 104193988 A CN104193988 A CN 104193988A CN 201410441173 A CN201410441173 A CN 201410441173A CN 104193988 A CN104193988 A CN 104193988A
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Abstract
The invention relates to a method for flocculating and sterilizing a fermentation solution of epsilon-polylysine, belonging to the field of biochemical engineering. The method comprises the steps of regulating the pH value of the fermentation solution to 0.5-4.0; slowly adding a 20-1200mg/L flocculant under a stirring condition, and slowly stirring for 10-20min; and adding 0.5-1.0% of filter aid, and then, pressing the flocculated fermentation solution into a plate and frame filter press to filter. Compared with a separation way which is commonly used at present, the method disclosed by the invention has the advantages that heating and diluting are not needed, the solid-liquid separation speed is high, the clarity of filtrate is high; the equipment investment and the maintenance cost are low; the yield of the epsilon-polylysine and the removal rate of hybrid protein are high; and the method is suitable for large-scale industrial production.
Description
Technical field
The present invention relates to a kind of method of epsilon-polylysine fermented liquid Flocculation, particularly a kind of method of removing thalline, purifying epsilon-polylysine in epsilon-polylysine fermented liquid with flocculation agent, belongs to biological chemical field.
Background technology
Epsilon-polylysine (ε-poly-L-lysine, ε-PL) is a kind of homotype aminoacid polymers being produced by microorganism secretions such as streptomycete, filamentous fungus or genus bacillus.It generally passes through α-COOH and ε-NH by 25-35 1B monomer
2dehydrating condensation forms, and molecular weight is generally 2500-4500Da.There is the features such as antimicrobial spectrum is wide, good water solubility, thermostability is strong, security is high due to ε-PL, be widely used in the foodstuffs industry of the countries and regions such as Japan, Korea S, America and Europe at present mainly as biological food sanitas.In addition, ε-PL is also used to the applied research in the field such as pharmaceutical carrier, gene chip.Therefore, ε-PL is considered to a kind of infant industry biological technology products with essential industry using value, is subject in recent years increasing food and technical field of biological material and pays close attention to.The production of ε-PL at present can only realize in a few countries, mainly concentrates on Japan.But, along with the steady lifting of domestic ε-PL fermentation level, possess the potentiality of industrialization.Therefore, research and development ε-PL high efficiency separation and extractive technique are most important to realizing its suitability for industrialized production.
ε-PL generally can only produce by microbe fermentation method at present.ε-PL fermented liquid is a very complicated system, except containing a small amount of target product (3-4%, dry weight) outside, also contain a large amount of somatic cells (20-30%, wet thallus volume), cell debris, biomacromolecule (as protein and nucleic acid) and colloidalmaterial etc.Meanwhile, fermented liquid also has higher apparent viscosity and presents typical non-Newton fluid characteristic; In addition, somatic cells is suitable with fermented liquid density, has formed stable colloidal solution.Therefore the solid-liquid separation that, uses general separation method to carry out ε-PL fermented liquid is very difficult.But if will realize ε-PL high efficiency separation and extraction, first remove the solid such as thalline and biomacromolecule and the fixing impurity of solubility in fermented liquid and be needs the problem that solves.
The method that separates and remove about somatic cells in ε-PL fermented liquid at present mainly contains centrifuging and filtration method.But in industrial production, centrifuging exists that facility investment is large, working cost is high, the not high shortcomings of separation efficiency; In addition, when centrifuging separates for the viscosity fermented liquid higher with solid content, also need to carry out dilution process, so just additionally increased the consumption of water resources and the load of subsequent extracted operation.Also have a kind of filter method based on acidifying and thermally denature to carry out solid-liquid separation processing to ε-PL fermented liquid, but, heating operation tends to produce a large amount of pigments, certainly will increase burden to follow-up bleaching process, also easily reduces yield and the purity of product and affects finished product outward appearance.In addition, somatic cells, in the situation that not forming flocs unit, can form compressible glue-like filter cake on filter cloth surface when filtration, very easily stops up filter cloth, causes filtering rate to decline rapidly, and time consumption and energy consumption and efficiency are low.
Summary of the invention
The object of the invention is to overcome above-mentioned weak point, provide a kind of do not need heating and dilution process and can high efficiency separation epsilon-polylysine fermented liquid in the method for thalline, reach the object of ε-PL fermented liquid being carried out to effective solid-liquid separation.
The inventive method comprises the following steps: ε-PL fermented liquid pH value is adjusted to pH0.5-4.0, slowly adds 20-1200mg/L flocculation agent under agitation condition.Slowly stir 10-20 minute.
In one embodiment, also, to add 0.5%-1.0% (g/100mL) flocculating aids in the fermented liquid of flocculation treatment, then the fermented liquid having flocculated is pressed in plate-and-frame filter press and is filtered.
Described ε-PL fermented liquid preference chain mould, filamentous fungus or genus bacillus ε-PL fermented liquid.Described streptomycete preferred white streptomycete.
Described pH value regulates preferred mineral acid.Described mineral acid comprises hydrochloric acid, sulfuric acid, phosphoric acid etc.
The preferred polymer organic anion of described flocculation agent flocculation agent, can be polyacrylic acid, polyacrylate, or both mixtures.Before flocculation agent uses, preferred dissolution is mixed with the aqueous solution of 0.1～2% (g/100mL).
The further optimization polypropylene acid of described flocculation agent sodium.
The effect of described flocculating aids is to reduce cake filtration resistance.Preferably diatomite.
Described flocculating aids is preferred perlite also.
Compared with the prior art the present invention has the following advantages:
1, cost is low: equipment one-time investment is low, and operation and maintenance cost is low; Do not need dilution, saved water resources, reduced subsequent extracted operational load yet; Do not need heating, reduced energy consumption yet.
2, high-level efficiency: fermented liquid is adjusted to extremely sour condition (pH0.5-4.0) by the present invention, makes thalline surface and the abundant cationization of protein can form obvious flocs unit in conjunction with anionic flocculant again, and can AUTOMATIC ZONING.Flocculation agent promoted somatic cells and/or biomacromolecule particle agglomerating, reduced fermentation broth viscosity, improved filtering rate, flux reaches 100L/h/m
2left and right.
3, the high product rate of recovery and albumen clearance: filtered liquid thalline is removed thoroughly, clarity is high; The epsilon-polylysine rate of recovery is greater than 95%; Protein removal rate reaches more than 70%.
Embodiment
Below in conjunction with embodiment, to being further described according to technical solution of the present invention:
Wherein, the measuring method of ε-PL adopts Itzhaki method (Itzhaki, R.F, Colorimetric method for estimating polylysine and polyarginine.Analytical Biochemistry, 1972:50,569 – 574); The measuring method of albumen adopts Bradford method (Bradford, M.M.A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.Analytical Biochemistry, 1976,72:248 – 254).
Embodiment 1
Fermented liquid is containing epsilon-polylysine 30g/L, dry cell weight 40g/L, pH3.8, protein concentration 1.2g/L.Utilize concentrated hydrochloric acid that fermented liquid pH value is adjusted to 0.5, the sodium polyacrylate that is 2% by concentration (molecular weight 5,000,000) solution slowly joins in the fermented liquid after above-mentioned acidifying, and be accompanied by mild stirring, until sodium polyacrylate final concentration reaches 800mg/L, then continue to stir 10 minutes; In fermented liquid, add 0.5% diatomite subsequently, stir.Above-mentioned fermented liquid is squeezed in plate-and-frame filter press, and pressure-controlling is filtered at 0.05MPa, and keeps constant pressure, finishes until filter.To be filteredly close on end, filter cake is rinsed with the water of 5% fermentating liquid volume, and filter cake is dried up with high-pressure air.Filtering omnidistance average filtration speed is 90L/h/m
2, ε-PL yield 95.5%, protein removal rate 74.4%.
Embodiment 2
Fermented liquid is containing epsilon-polylysine 40g/L, dry cell weight 60g/L, pH4.0, protein concentration 2.2g/L.Utilize the vitriol oil that fermented liquid pH value is adjusted to 2.0, the polyacrylic acid that is 0.5% by concentration (molecular weight 1,000 ten thousand) solution slowly joins in the fermented liquid after above-mentioned acidifying, and be accompanied by mild stirring, until sodium polyacrylate final concentration reaches 20mg/L, then continue to stir 20 minutes; In fermented liquid, add 0.5% perlite subsequently, stir.Above-mentioned fermented liquid is squeezed in plate-and-frame filter press, and pressure-controlling is filtered at 0.1MPa, and keeps constant pressure, finishes until filter.To be filteredly close on end, filter cake is rinsed with the water of 5% fermentating liquid volume, and filter cake is dried up with high-pressure air.Filtering omnidistance average filtration speed is 100L/h/m
2, ε-PL yield 96.3%, protein removal rate 73.2%.
Embodiment 3
Fermented liquid is containing epsilon-polylysine 20g/L, dry cell weight 30g/L, pH3.5, protein concentration 1.2g/L.Utilize phosphoric acid that fermented liquid pH value is adjusted to 3.0, the sodium polyacrylate that is 1.0% by concentration (molecular weight 1,000 ten thousand) solution slowly joins in the fermented liquid after above-mentioned acidifying, and be accompanied by mild stirring, until sodium polyacrylate final concentration reaches 1200mg/L, then continue to stir 5 minutes; In fermented liquid, add 0.5% diatomite subsequently, stir.Above-mentioned fermented liquid is squeezed in plate-and-frame filter press, and pressure-controlling is filtered at 0.15MPa, and keeps constant pressure, finishes until filter.To be filteredly close on end, filter cake is rinsed with the water of 5% fermentating liquid volume, and filter cake is dried up with high-pressure air.Filtering omnidistance average filtration speed is 120L/h/m
2, ε-PL yield 95.2%, protein removal rate 72.5%.
Embodiment 4
Fermented liquid is containing epsilon-polylysine 35g/L, dry cell weight 45g/L, pH4.0, protein concentration 1.6g/L.The sodium polyacrylate that is 1.5% by concentration (molecular weight 3,000 ten thousand) solution slowly joins in the fermented liquid after above-mentioned acidifying, and is accompanied by mild stirring, until sodium polyacrylate final concentration reaches 600mg/L, then continues to stir 5 minutes; In fermented liquid, add 1.0% diatomite subsequently, stir.Above-mentioned fermented liquid is squeezed in plate-and-frame filter press, and pressure-controlling is filtered at 0.15MPa, and keeps constant pressure, finishes until filter.To be filteredly close on end, filter cake is rinsed with the water of 5% fermentating liquid volume, and filter cake is dried up with high-pressure air.Filtering omnidistance average filtration speed is 110L/h/m
2, ε-PL yield 94.2%, protein removal rate 78.5%.
Embodiment 5
Fermented liquid is containing epsilon-polylysine 30g/L, dry cell weight 42g/L, pH0.5, protein concentration 1.5g/L.The sodium polyacrylate that is 1.5% by concentration (molecular weight 3,000 ten thousand) solution slowly joins in the fermented liquid after above-mentioned acidifying, and is accompanied by mild stirring, until sodium polyacrylate final concentration reaches 1000mg/L, then continues to stir 5 minutes; In fermented liquid, add 1.0% diatomite subsequently, stir.Above-mentioned fermented liquid is squeezed in plate-and-frame filter press, and pressure-controlling is filtered at 0.15MPa, and keeps constant pressure, finishes until filter.To be filteredly close on end, filter cake is rinsed with the water of 5% fermentating liquid volume, and filter cake is dried up with high-pressure air.Filtering omnidistance average filtration speed is 108L/h/m
2, ε-PL yield 92%, protein removal rate 78%.
Although the present invention with preferred embodiment openly as above; but it is not in order to limit the present invention, any person skilled in the art, without departing from the spirit and scope of the present invention; all can do various changes and modification, therefore protection scope of the present invention should be with being as the criterion that claims were defined.
Claims (10)
1. a method for epsilon-polylysine fermented liquid Flocculation, comprises the following steps: epsilon-polylysine fermented liquid pH value is adjusted to pH0.5-4.0; Under agitation condition, slowly add 20-1200mg/L flocculation agent.
2. method according to claim 1, is characterized in that, also comprises to add 0.5%-1.0% flocculating aids in the fermented liquid of flocculation treatment, then the fermented liquid having flocculated is pressed in plate-and-frame filter press and is filtered.
3. method according to claim 1, is characterized in that, described pH value regulates and adopts mineral acid.
4. method according to claim 3, is characterized in that, described mineral acid is hydrochloric acid, sulfuric acid or phosphoric acid.
5. method according to claim 1, is characterized in that, described flocculation agent is polymer organic anion flocculation agent.
6. method according to claim 5, is characterized in that, described flocculation agent is polyacrylic acid or polyacrylate or both mixtures.
7. method according to claim 5, is characterized in that, described flocculation agent is sodium polyacrylate.
8. method according to claim 2, is characterized in that, the preferred diatomite of described flocculating aids.
9. method according to claim 2, is characterized in that, the preferred perlite of described flocculating aids.
10. method according to claim 1, is characterized in that, described ε-PL fermented liquid is streptomycete, filamentous fungus or genus bacillus ε-PL fermented liquid.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106380592A (en) * | 2016-11-04 | 2017-02-08 | 江南大学 | Method for extracting epsilon-polylysine and hydrochloride thereof from fermentation liquid |
| CN111153478A (en) * | 2020-01-19 | 2020-05-15 | 南昌诺汇医药科技有限公司 | Polylysine fermentation liquid processing apparatus |
| CN111533652A (en) * | 2020-05-25 | 2020-08-14 | 吉林中粮生化有限公司 | Method and system for separating lactate |
| CN115746067A (en) * | 2022-11-15 | 2023-03-07 | 武汉科诺生物科技股份有限公司 | Solid-liquid separation method of fermentation liquor |
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| CN102363797A (en) * | 2011-10-28 | 2012-02-29 | 天津科技大学 | Method for producing epsilon-poly-L-lysine |
| CN102703407A (en) * | 2012-06-18 | 2012-10-03 | 江南大学 | Method for preparing leucine aminopeptidase through fermentation of bacillus subtilis engineering bacteria |
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| CN1696279A (en) * | 2005-02-04 | 2005-11-16 | 南京工业大学 | Method for preparing epsi-polylysine and its salt by using Kitasatosporia PL6-3 |
| CN101701069A (en) * | 2009-09-22 | 2010-05-05 | 浙江银象生物工程有限公司 | Method for extracting epsilon-polylysine and salt thereof |
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Cited By (4)
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| CN106380592A (en) * | 2016-11-04 | 2017-02-08 | 江南大学 | Method for extracting epsilon-polylysine and hydrochloride thereof from fermentation liquid |
| CN111153478A (en) * | 2020-01-19 | 2020-05-15 | 南昌诺汇医药科技有限公司 | Polylysine fermentation liquid processing apparatus |
| CN111533652A (en) * | 2020-05-25 | 2020-08-14 | 吉林中粮生化有限公司 | Method and system for separating lactate |
| CN115746067A (en) * | 2022-11-15 | 2023-03-07 | 武汉科诺生物科技股份有限公司 | Solid-liquid separation method of fermentation liquor |
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